CN106103427B - Indazole compound and preparation method thereof - Google Patents

Indazole compound and preparation method thereof Download PDF

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CN106103427B
CN106103427B CN201480048261.9A CN201480048261A CN106103427B CN 106103427 B CN106103427 B CN 106103427B CN 201480048261 A CN201480048261 A CN 201480048261A CN 106103427 B CN106103427 B CN 106103427B
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indazole
compound
formula
imidazoles
chloro
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CN106103427A (en
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杜姆巴拉·斯里尼瓦萨·雷迪
柴塔尼亚·萨克塞纳
卡希纳特·科米里谢特利
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Santanyi Protein Analysis Co Ltd
Council of Scientific and Industrial Research CSIR
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Council of Scientific and Industrial Research CSIR
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
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    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/416—1,2-Diazoles condensed with carbocyclic ring systems, e.g. indazole
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    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
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    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond

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Abstract

This patent discloses the indazole compounds and the like of new general formula 1, are used to treat the purposes of diabetes, diabetic complication, cardiovascular functional disorder or related disease, include their pharmaceutical composition and preparation method thereof.

Description

Indazole compound and preparation method thereof
Technical field
The present invention relates to new indazole compounds of general formula 1 and preparation method thereof.The present invention is more particularly directed to new indazoles Close object, its derivative and its synthetic method.The invention further relates to the new indazoles of general formula 1 for treating diabetes, diabetes simultaneously Send out disease, metabolic disorder, cardiovascular functional disorder (including hypertension), autoimmunity and the relevant illness of inflammation or in fatty acid and There are glucose processing impaired (impaired glucose disposal) and energy consumption imbalances between glucose mechanism The purposes of disease, and further relate to the pharmaceutical composition comprising them.
Background technique and the prior art
Diabetes B (Type 2 diabetes mellitus, T2DM) is most common, chronic and life-threatening disease One of.Every year, the illness rate of T2DM is continuously improved in worldwide, and recently, the World Health Organization (World Health Organization, WHO) prediction, to the year two thousand thirty, the patient numbers for being diagnosed as T2DM will be more than 3.66 hundred million.T2DM Clinical symptoms be due to hypoinsulinism, insulin resistance or both caused by blood glucose level raising.
2004/0009976 A1 of US discloses the compound of formula (A1):
And its for treating type-2 diabetes mellitus and the purposes that stimulates insulin secretion in mammals.
2003/0109550 A1 of US discloses the compound of formula A2,
Wherein, B is 5 or 6 yuan of saturations or unsaturated heterocycle, wherein the heterocycle is optionally by R1、R2Replace with R12；X choosing From N and C；Y and Z is independently selected from N, CH, CR3, S and O；R3The amidine that is selected from: being substituted or be unsubstituted, alkyl amino, amino Alkyl, CONHR7、 NH2、NHCOR6And CH2NHCOR6。
0418845 B1 of EP discloses new pyrazole derivatives, preparation method and the medicine comprising the pyrazole derivatives Compositions.It provides the compound of formula A3:
Wherein, R1 is the aryl that can be replaced by substituent group selected from the following: 40 low alkyl groups, halogen, lower alkoxy, low It is grade alkylthio group, lower alkyl sulfinyl, lower alkylsulfonyl radicals, hydroxyl, lower alkylsulfonyloxy, nitro, amino, rudimentary Alkyl amino, acylamino- and low alkyl group-(acyl group) amino；Or heterocycle；R2 is hydrogen；By amino, low-grade alkyl amino, halogen Or the methyl that acyloxy replaces；Acyl group；45 acylamino-s；Cyano；Halogen；Lower alkylthio；Lower alkyl sulfinyl；Or heterocycle Base；And R3 is the aryl replaced by following substituent group: low alkyl group, lower alkylthio, lower alkyl sulfinyl, halogen, Amino, low-grade alkyl amino, acylamino-, low alkyl group (acyl group) amino, lower alkoxy, cyano, hydroxyl or acyl group；Either The heterocycle that can be replaced by lower alkylthio, lower alkyl sulfinyl or 50 lower alkylsulfonyl radicals.
US 4,436,913 discloses 1H- indazole and 2H- indazole derivative and its acid-addition salts and containing these drop blood The 1H- indazole of the formula A4 of pressure and the drug of 2H indazole derivative and its acid-addition salts,
Wherein R1 can be located at 1 or 2 on the nitrogen-atoms in Formulas I.Group R1, R2 and R3 represent hydrogen or common rudimentary Molecular radical.R group is 2- imidazolinylamino or 3, and 4,5,6- tetrahydro-pyrimidine base amino, wherein these groups can also be with it Tautomeric form exists.These groups can also be in the aryl in R1 group, and in this case, R group can also be Halogen atom.When R1 group is simultaneously aryl or aralkyl, R can only represent miscellaneous in 4 or 7 of 1H indazole derivative One of ring secondary amine or tertiary amine groups.
6,878,735 B2 of US discloses the imidazoline of formula A5,
Wherein R1 is optionally substituted aryl, and R2 is selected from: alkyl, acyl group, aryl, aralkyl, containing 5 to 14 rings at The heteroaryl and heterocycle containing 5 to 12 ring members of member；R3 is optionally substituted aryl, and R4 is optionally substituted aralkyl Base, and R5 is selected from hydrogen and alkyl, and all these is optionally substituted.
7,541,376 B2 of US provides the new 1H- indazole compound with excellent JNK inhibiting effect.More specifically Ground, it provides compound shown in formula A6, its salt or its hydrate,
Wherein R1It is C6-C14Aromatic cyclic hydrocarbon radical etc.；R2、R4And R5Each independently represent hydrogen atom, halogen atom, cyano Deng；L is singly-bound or C1-C6Alkylidene etc.；X is group shown in singly-bound or-CO-NH- or-NH-CO- etc.；Y is C3-C8Cycloalkanes Base, C6-C14Aromatic cyclic hydrocarbon radical or 5 yuan are to 14 yuan of aromatic heterocyclic radicals etc..
2011/0034441 A1 of US discloses the compound of formula A7,
Wherein R1、R2、R4、R5、R6、R7And R8Independently selected from: H, C1-9Alkyl, halogen ,-CF3、-(C1-9Alkyl)nCarbocyclic ring Base R12、-(C1-9Alkyl)nHeterocycle R12、-(C1-9Alkyl)nAryl R12、-(C1-9Alkyl)nHeteroaryl R12、-(C1-9Alkyl)nOR9、-(C1-9Alkyl)nSR9、-(C1-9Alkyl)nS (=O) R10、-(C1-9Alkyl)nSO2R9、-(C1-9Alkyl)nN(R9) S (=O) R10、-(C1-9Alkyl)nN(R9)SO2R9、-(C1-9Alkyl)nSO2N(R9)2、-(C1-9Alkyl)nN(R9)2、-(C1-9Alkyl)nN(R9)C (=A) N (R9)2、-(C1-9Alkyl)nNR9C (=O) OR9、-(C1-9Alkyl)nC (=A) N (R9)2、-(C1-9Alkyl)nN(R9) C (= A)R9、-(C1-9Alkyl)nOC (=O) N (R9)2、 -NO2、-CN、-(C1-9Alkyl)nCO2R9With-(C1-9Alkyl)nC (=A) R9。
2002/0161022 A1 of US discloses the compound of formula A8,
Wherein R1 is the alkyl for being substituted or being unsubstituted, aryl, heteroaryl, carbocyclic ring, heterocycle etc.；R2It is to be substituted or not Alkyl, aryl, heteroaryl, carbocyclic ring, heterocycle for being substituted etc..
WO2011057959 discloses the compound or pharmaceutically acceptable salt thereof of formula A9,
Wherein, R1, R2, R3 are hydrogen, halogen, low alkyl group or alkoxy independently of one another；R4 is hydrogen, is unsubstituted Low alkyl group, or by one to four independently selected from low alkyl group replaced methyl, (=O) and the substituent group of-COOH；X is CH or N；And Y is hydrogen or-NH2.In addition, it is related to compound A9 or its officinal salt for treating or preventing metabolic disease With the purposes of illness.
In 20 (2003) 201-208 of European Journal of Pharmaceutical Sciences, Entitled " 2- (4,5-Dihydro-1H-imidazol-2-yl) indazole (indazim) of F.Saczewski etc. The article of derivatives as selective I2 imidazoline receptor ligands " reports one system of synthesis Arrange different substituted 2- (4,5- dihydro -1H- imidazoles -2- base) indazoles and 2- (4,5- dihydro -1H- imidazoles -2- base) -4,5,6,7- Tetrahydrochysene indazole.In addition, which reports the chloro- 2- of 4- (4,5- dihydro -1H- imidazoles -2- base) indazoles (3f, 4-Cl-indazim) to exist Good affinity is shown on imidazoline I2 receptor and between such imidazoline ligand in α -2- adrenaline Unprecedented low-affinity is shown on receptor.
ChemCommun(Camb).2010Nov 28；46 (44): in 8407-9, entitled " Click " of YuhuiLoh synthesis of small molecule-peptide conjugates for organelle-specific The article report of delivery and inhibition of lysosomal cysteine proteases " is small for synthesizing Molecule inhibitor-peptide conjugate with realize organelle-specificity delivering click chemistry method.It also offers a kind of methods, wherein Using N-methylmorpholine (N-methylmorpholine, NMM) isobutyl chlorocarbonate (isobutyl chloroformate, ISCF) and COOH group is converted CONNH2ME by methyl hydrazonium salt.
Organic Mechanisms, in page 2010,321 to 338 article " Carboxylic Compounds, Nitriles, and Their Interconversion " are reported with forming trifluoroacetic acid F3C-CO2The reagent of H is through trifluoro second Acid anhydrides mediates neopentanoic acid amide (A) to be dehydrated into neopentanoic acid nitrile (B).
In Bioorganic&Medicinal Chemistry 15 (2007) 6782-6795, Rajesh H.Bahekar, Mukul R.Jain、Pradip A.Jadav、Vijay M.Prajapati、Dipam N.Patel、Arun A.Gupta、Ajay Sharma, Robby Tom, Debdutta Bandyopadhya, Honey Modi and Pankaj R.Patel it is entitled " Synthesis and antidiabetic activity of 2,5-disubstituted-3-imidazol-2-yl- The article of pyrrolo [2,3-b] pyridines and thieno [2,3-b] pyridines " is reported to be used at 120 DEG C Ethylenediamine, P2S5Imidazoline was converted by nitrile through 5 hours.
The definition (Trends in Pharm.Sci.21,259-265 2000) of pathogenic effects based on insulin resistance, The drug used in treating diabetes belongs to following therapeutic categories: insulin, sulfonylurea, melbine, alpha-glucosidase Inhibitor (acarbose) and thiazolidinediones (troglitazone).Insulin is most well-known drug, and it is considered as Reference drug in treating diabetes.But insulin therapy has the disadvantages that drug is only applied by parenteral route, Blood glucose level must constantly be controlled, it may occur that local allergy, insulin resistance are forced with the time meaningfully Drug dose is improved, local tolerance is poor.
In addition, other treatment methods are not without disadvantage, it is sometimes even more significant.Such as sulfonylurea, individually apply With or with insulin or with other oral hypoglycaemic drugs be administered in combination when can cause hypoglycemia.Be used alone or with sulfonylureas group It closes the melbine used and avoids use when having nephrosis and liver disease, and lactic acidosis state can be induced.Acarbose individually makes For reducing level of postprandial blood sugar when being applied in combination with or with sulfonylureas, but it usually induces secondary make in gastronintestinal system level With.The troglitazone only used with insulins combinations can induce hepatotoxic effect.
Therefore, there is an urgent need to develop some new normal physiological responses that can attempt to retain to meal ingestion for blood glucose The treatment method of control.Method as a kind of is the exploitation based on Insulin secretagogues (secretagogue), the pancreas Island element succagoga will not cause glucose production in the case where basal plasma glucose is horizontal, but only show that glucose dependent insulin is released It puts.
In view of problems of the prior art and to novel drugs, (it shows that glucose dependent insulin is released Put, overcome the side effect for the treatment of diabetes or diabetes-related complication) long-awaited demand, inventor has been proposed The present invention.The invention discloses new indazole compound, its derivative and its synthetic methods.The compound is to 1 type and 2 types sugar Urine disease and related complication (such as diabetic neuropathy, diabetic retinopathy and due to glucose blood level continued jitters Caused by a variety of vascular diseases) show anti-diabetic activity.
Goal of the invention
The main object of the present invention is to provide the new indazole compound of general formula 1.
It is a further object to provide the methods of synthesis 1 compound of formula.
A further object of the present invention is to provide for treating diabetes, diabetes-related complication and the chemical combination of hypertension The pharmaceutical composition of object.
A further object of the present invention is to provide the new indazole of general formula 1 for treating diabetes, diabetic complication, generation Thank disorder, cardiovascular functional disorder (including hypertension), autoimmunity and the relevant illness of inflammation or in fatty acid and grape There are the impaired purposes with the disease of energy consumption imbalance of glucose processing between sugared approach and relevant triglyceride levels.
Summary of the invention
Therefore, the present invention provides the indazole compound of formula 1,
Wherein:
R1It is hydrogen or alkyl or aryl or heteroaryl；
R2It is H or halogen；
R3、R4、R5And R6It independently is hydrogen or alkyl, aryl, heteroaryl；Selected from R3、R4、 R5And R6Any two it is adjacent In addition group, which can be formed, can contain heteroatomic 3 to 8 member ring；And the like, position isomer, stereoisomer, derivative And officinal salt.
In one embodiment of the invention, the indazole compound of formula 1 is indicated by following compounds:
A.5- chloro- 3- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
B.5- chloro- 3- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl -3a, 7a- dihydro -1H- indazole；
C.5- chloro- 3- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole；
D.5- chloro- 3- (4,5- dihydro -1H- imidazoles -2- base) -1- ethyl -1H- indazole；
E.1- the chloro- 3- of benzyl -5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
F.3- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
G.3- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl-1 H- indazole；
H.3- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole.
In one embodiment of the invention, the preparation method of the indazole compound of formula, wherein the method includes with Lower step:
I. 5- chlorine indazole 3- carboxylic acid (2) is converted by 5- chlorisatide (5-chloro isatin) (1)；
Ii. under argon gas, compound 5- chlorine indazole 3- carboxylic acid (2) are handled with isobutyl chlorocarbonate and N-methylmorpholine, with After react with ammonia water to obtain the chloro- 1H- indazole -3- formamide (3) of 5-；
Iii. cyanalation to obtain with pyridine and the trifluoroacetic anhydride processing chloro- 1H- indazole -3- formamide (3) of compound 5- It closes object (4)；
Iv. in the solvent selected from acetone, react cyano compound (4) to be substituted with potassium carbonate and alkyl halide Indazole formonitrile HCN；
V. in P2S5In the presence of, make the indazole formonitrile HCN being substituted and diamine reactant to obtain 1 compound of general formula.
In another embodiment of the present invention, alkyl halide used in step (iv) be selected from bromic ether, methyl iodide, Benzyl bromide.
In another embodiment of the present invention, diamines used in step (v) is selected from 1,2- cyclohexanediamine, second Diamines.
In another embodiment of the present invention, pharmaceutical preparation, it includes as active constituent 1 compound of formula or Its analog, position isomer, stereoisomer, derivative and officinal salt, and it is one or more of pharmaceutical acceptable carrier, dilute Release agent or excipient.
In another embodiment of the present invention, the indazole compound of formula 1 can be used for treating diabetes, diabetes complicated Disease, metabolic disorder, cardiovascular functional disorder or in which that there are glucose processing is impaired, triglyceride levels change or beta cell function The related disease that can be reduced.
In another embodiment of the present invention, the indazole compound of formula 1, wherein it is changed with the formula 1 of therapeutically effective amount It closes object or its analog, position isomer, stereoisomer, derivative and officinal salt is applied to mammal.
Brief description
Fig. 1: the insulin secreting ability of the NDS100178 and NDS100179 in different three days is described.
Fig. 2: the glucose dependency activity of NDS100178 and NDS100179 is described.
Fig. 3: the insulinotropic activity of NDS100178 at low concentrations and NDS100179 are described.
Fig. 4: a) oral glucose tolerance test is described: blood glucose in the presence of NDS100179 in C57BL mouse Reduction；B) oral glucose tolerance test: NDS100179 dose dependent % in total AUC (glucose) in C57BL mouse Variation.
Fig. 5: it has retouched GSIS in a) people's pancreas islet and has improved；B) GSIS is improved in C57BL mouse；C) GSIS is mentioned in MIN6 cell It is high.
Fig. 6: the glycemic control in db/db mouse a) handled through NDS100179 is described；B) it is handled through NDS100179 Db/db mouse in glucose tolerance；C) control or through NDS100179 handle db/db mouse in measured body weight.
Fig. 7: the triglycerides measurement in control or the db/db mouse handled through NDS100179；B) it compares or passes through Instreptozotocin Induced measurement in the db/db mouse of NDS100179 processing.
Fig. 8: the glucose uptake in HepG2 cell is described.
Fig. 9: the PK of NDS100179 in C57BL/6JRccHsd mouse is described.
Figure 10: the oral glucose tolerance test in C57BL/6JRccHsd mouse is described.
Detailed description of the invention
The present invention relates to new indazole compounds of general formula 1 and preparation method thereof.The present invention provides the indazoles of general formula 1 Close noval chemical compound of object and the like, position isomer, stereoisomer, derivative and officinal salt and the like, position Set isomers, stereoisomer, derivative and its officinal salt.
Wherein:
R1It is hydrogen or alkyl or aryl or heteroaryl；
R2It is H or halogen；
R3、R4、R5And R6It independently is hydrogen or alkyl, aryl, heteroaryl；
Selected from R3、R4、R5And R6Any two adjacent group can be formed and can in addition contain heteroatomic 3 to 8 member ring.
In a preferred embodiment, the present invention provides 1 compounds of formula selected from the following:
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (NDS100178)；
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl -3a, 7a- dihydro -1H- indazole (7, NDS100179)；
The chloro- 3- of 5- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole (8, NDS100281)；
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- ethyl -1H- indazole (10, NDS100282)；
The chloro- 3- of 1- benzyl -5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (12, NDS100283)；
3- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (16, NDS100277)；
3- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl-1 H- indazole 18 (NDS100278)；
3- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole (19, NDS100279).
In another embodiment, the present invention provides the noval chemical compounds that the isatin compound by Formula II prepares general formula 1 Method,
Wherein R2It is H or halogen；
The following steps are included:
Carboxylic acid is converted by the isatin compound of formula (II)；
With the carboxylic acid compound of isobutyl chlorocarbonate and N-methylmorpholine processing step (a), then react with ammonia water to obtain To amide；
With pyridine and the amide compound of trifluoroacetic anhydride processing step (b) to provide cyano compound；
In P2S5In the presence of, make cyano compound and the diamine reactant of step (c) to obtain desired formula (1) compound.
In a preferred embodiment, the present invention provides the method for the noval chemical compound of the formula that is used to prepare 5, packets It includes:
5- chlorine indazole 3- carboxylic acid (2) is converted by 5- chlorisatide (1)；
Amide (3) are converted by the compound (2) of step (a)；
With pyridine and the amide of trifluoroacetic anhydride processing step (b) 3 to provide cyano compound (4)；
In P2S5In the presence of, make compound (4) and the reacting ethylenediamine of step (c) to obtain compound (5).
The preparation method of compound 5 is described in scheme 1.
Scheme 1:
Scheme: 1
In another preferred embodiment, described the present invention provides the method for the noval chemical compound of the formula that is used to prepare 7 Method includes:
5- chlorine indazole 3- carboxylic acid (2) is converted by 5- chlorisatide (1)；
Amide (3) are converted by the compound (2) of step (a)；
With pyridine and the amide of trifluoroacetic anhydride processing step (b) 3 to provide cyano compound (4)；
The compound (4) of step (c) is methylated to obtain compound (6),
In P2S5In the presence of, make compound (6) and the reacting ethylenediamine of step (c) to obtain compound (7).
The method for being used to prepare compound 7 is described in scheme 2:
(scheme: 2)
Method disclosed in the present invention has high yield and selectivity, and is commericially feasible.
In one aspect of the invention, 1 compound of formula can have to 1 type and diabetes B and related complication (such as sugar Urinate characteristic of disease neuropathy, diabetic retinopathy and a variety of angiocarpy diseases as caused by blood glucose and triglyceride levels continued jitters Disease) anti-diabetic activity.
In one aspect of the invention, the insulinotropic activity of 1 compound of formula is had studied.1 compound of formula is in 0.1 μ g/ml Insulinotropic activity is shown in the range of to 10 μ g/ml.1 compound of formula shows concentration dependent insulinotropic activity.
On the other hand, by the prior art chemical combination of the insulinotropic activity of 1 compound of formula and US 20040009976 The activity of object compares, and by the results are shown in Table 1.Compared with those disclosed in the prior art, 1 compound of formula shows to increase Strong insulinotropic activity.
In yet another aspect, the present inventor has studied the insulin secretion of concentration of glucose dependence or Portugal in a variety of models The insulin secretion (GSIS) of grape sugar stimulation.This research has been carried out in people's pancreas islet, C57BL mouse and MIN6 cell.Formula 1 Compound induces GSIS, and is evident that this point by reference to Fig. 5.Test also demonstrates, the GSIS of the compound of the present invention Effect is independent of biosystem and its composition.
In addition, under study for action, in the OGTT test in diabetes animal model (db/db mouse), when with 1 compound of formula At processing animal 30 days, the compound provides excellent control to the fasting blood sugar level gradually increased and improves grape Sugared tolerance.This provides the direct instruction (Fig. 6 a and 6b) of the anti-diabetic characteristic of 1 compound of formula.In addition, through compound Preferably retain in the animal of processing/restore Instreptozotocin Induced, it was confirmed that the additional benefit (Fig. 6 c) of the compound.
In another discovery, in diabetes animal model (db/db mouse), when with 1 compound of formula handle animal 30 It when, the compound reduces triglyceride levels (Fig. 7).Low triglyceride levels are related to improved health of heart simultaneously And compound provides the benefit of significant treatment cardiovascular and cerebrovascular.
With reference to Fig. 8, the glucose uptake of the HepG2 cell in the presence of compound is had studied.The Portugal of HepG2 cell Grape Sugar intake represents the model of the glucose uptake of research liver cell.The glucose uptake of raising is characterized in that improved pancreas islet Plain sensibility, and glucose uptake can be stimulated and improve the compound of insulin sensitivity and can be used for the control of diabetes B, Because they are finally beneficial to maintain glucose homeostasis come the glucose of self-circulation system by reducing., it is apparent that changing It closes object and improves the glucose uptake in HepG2 cell, therefore indicate that compound can help to maintain glucose homeostasis and control 2 Patients with type Ⅰ DM.
In yet another aspect, present inventors have proposed the mechanisms of action of 1 compound of formula.By imidazoline receptor body running, change It closes object and improves the diglyceride in cell and the arachidonic acid in downstream.Arachidonic acid metabolite participates in insulin exocytosis and makees With.The new discovery of an inhibition LTA4H has had been established in the present inventor, can maintain arachidonic acid library in cell, pancreas can be improved The enzyme of island element secretion.NDS100178 inhibits LTA4H (IC50 < 500nM), and for the change from patent US20040009976 Close the IC50 of the LTA4H target of the embodiment 6 of object 5- chloro-2-methyl -3- (4,5- dihydro -1-H- imidazoles -2- base) -1H- indoles Greater than 10uM.
In yet another embodiment, pharmaceutical composition is provided, it includes the compound or stereoisomer of formula (I), Or ester or its officinal salt and pharmaceutical acceptable carrier, diluent or excipient.
Pharmaceutical composition of the invention can be by by the compound of the present invention and pharmaceutical acceptable carrier appropriate, diluent or tax Shape agent is combined to prepare, and can be configured to the preparation of solid, semisolid, liquid or gas form, such as tablet, capsule Agent, powder, granule, ointment, solution, injection, gelling agent and microspheres agent (microsphere).
In a further embodiment, the present invention relates to " this hairs to the object application " effective quantity " with the disease Bright composition ".Therefore, any amount, any type of pharmaceutical composition can be used to apply by any effective treatment disease 1 compound of formula, the pharmaceutical composition containing 1 compound of formula are applied with approach.Apply the typical case of this pharmaceutical composition Approach includes but is not limited to: oral, through surface, percutaneous, sucking, parenteral, sublingual, mouth containing, per rectum, Via vagina and intranasally.
When preparing pharmaceutical composition of the invention so as to apply the composition to patient, the active constituent being included in is Bioavailable.The composition applied to object or patient can be taken to the form of one or more dosage units.Dosage form It can also be prepared as lasting, control, modification and instant dosage form.
In a word, it is believed that: 1 compound of formula makes the weight of itself and object by induction GSIS, control blood sugar concentration and applies Concentration is proportional, reduces triglycerides and recovery/reservation Instreptozotocin Induced to improve cardiovascular safety characteristic, is glycosuria Patient and object with its complication provide new alternative.
Embodiment below be provided and illustrating work of the invention in actual implementation, and be not necessarily to be construed as with Any mode limits the scope of the invention.
Embodiment
Embodiment: 1
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (NDS100178):
The chloro- 1H- indazole -3- formamide (3) of 5-:
Under argon gas at 0 DEG C, to 5- chlorine indazole 3- carboxylic acid1In anhydrous THF (20mL) solution of 2 (0.8g, 4.0mmol) Isobutyl chlorocarbonate (0.64mL, 4.9mmol) and N- methyl morpholine (0.7mL, 6.1mmol) are added, and stirs the mixture for 2 Hour.Then the NH of 10mL is added into the mixture3Aqueous solution, and mixture is stirred 1 hour at 25 DEG C.It is depressurizing Lower removing THF, obtains solid by filtered on buchner funnel, uses ether washing solid and is dried under vacuum to obtain as pale yellow The title compound 3 (0.5g, 63%) of color solid.IRυmax(film): cm-12925,2854,1463；
1H NMR (200MHz, DMSO-d6): δ 8.14 (d, J=1.6 Hz, 1H), 7.80 (bs, 1H), 7.66 (d, J= 8.9Hz, 1H), 7.45-7.39 (m, 2H)；MS:218 (M+Na)+.
The chloro- 1H- indazole -3- formonitrile HCN (4) of 5-:
The chloro- 1H- indazole -3- formamide 3 (0.60g, 3.0mmol) of 5- is dissolved in pyridine (6mL) and anhydrous methylene chloride In (6mL).It adds trifluoroacetic anhydride (1.0mL, 7.7mmol), and reaction is stirred 10 minutes at 25 DEG C.By reaction mixture It is concentrated in a vacuum, and residue is placed in ethyl acetate, then use water, saturated sodium bicarbonate and salt water washing.It will be organic It is mutually dried over sodium sulfate, filters and the chloro- 1H- indazole -3- formonitrile HCN of title compound 5- to obtain as light yellow solid is concentrated 4 (0.5g, 92%).IRυmax(film): cm-12233；
1H NMR (200MHz, CDCl3): δ 8.03 (d, J=9.1Hz, 1H), 7.96 (d, J=1.8 Hz, 1H), 7.75 (dd, J=9.1,1.8Hz, 1H)
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (5, NDS100178):
By the chloro- 1H- indazole -3- formonitrile HCN 4 (0.2g, 1.1mmol) of 5-, ethylenediamine (EDA, 4mL) and P2S5(0.1g, Mixture 0.4mmol) heats 5 hours at 120 DEG C.Reaction mixture is poured into and obtains solid in ice, passes through Buchner funnel Filtering, is washed, and the chloro- 3- of 5- (4, the 5- dihydro -1H- imidazoles-to obtain as pale solid are dried under vacuum with ether 2- yl) -1H- indazole compound 5NDS100178 (0.12g, 49%).Mp=248-249 DEG C；
1H NMR (200 MHz, DMSO-d6): δ 8.19 (s, 1H), 7.63 (d, J=8.9Hz, 1H), 7.36 (d, J= 8.9Hz, 1H), 3.66 (bs, 4H)；MS:221 (M+H)+.
Embodiment: 2
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -2- methyl -2H- indazole (NDS100179)
The chloro- 1- methyl-1 H- indazole -3- formonitrile HCN (6) of 5-:
By potassium carbonate (0.23g；1.6mmol) and methyl iodide (0.1mL；1.6mmol) it is added to the chloro- 1H- indazole -3- first of 5- 4 (0.1g of nitrile；0.5mmol) in the suspension in acetone (10mL).Reaction mixture is heated 4 hours at 65 DEG C, it is cooling And it filters.Filtrate is concentrated under reduced pressure, and is purified by column chromatography (silica gel 100-200 mesh, 3: 7 ethyl acetate: petroleum ether) The chloro- 1- methyl-1 H- indazole -3- formonitrile HCN 6 (0.8g, 75%) of 5- to obtain as white solid.Mp=154-155 DEG C；
1H NMR (200MHz, DMSO-d6): δ 8.03 (d, J=1.9 Hz, 1H), 7.96 (d, J=8.9Hz, 1H), 7.63 (dd, J=8.9,1.9Hz, 1H), 4.20 (s, 3H)
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl -3a, 7a- dihydro -1H- indazole (7, NDS100179):
By the chloro- 1- methyl-1 H- indazole -3- formonitrile HCN 6 (0.1g, 1.0mmol) of 5-, EDA (4mL) and P2S5(0.046g, Mixture 0.4mmol) heats 5 hours at 120 DEG C.Reaction mixture is poured into and obtains solid in ice, passes through Buchner funnel Filtering, washed with ether, and be dried under vacuum to obtain as white solid compound 7 (NDS100179) (0.060g, 50%).Mp=174-175 DEG C；
1H NMR (200MHz, DMSO-d6): δ 8.21 (d, J=2.0Hz, 1H), 7.77 (d, J=8.8Hz, 1H), 7.47 (dd, J=8.8,2.0Hz, 1H), 6.84 (bs, 1H), 4.11 (s, 3H), 3.62 (bs, 4H)13C NMR (100MHz, DMSO- d6) δ=160.1,139.6,132.0,128.2,127.2,122.4,120.1,111.2,35.2；MS；235(M+H)+.
Embodiment: 3
The chloro- 3- of 5- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole 8 (NDS100281):
By the chloro- 1H- indazole -3- formonitrile HCN 4 (0.1g, 0.5mmol) of 5-, 1,2- cyclohexane diamine (2mL) and P2S5(0.05g, Mixture 0.2mmol) heats 5 hours at 120 DEG C.Reaction mixture is poured into and obtains solid in ice, passes through Buchner funnel Filtering, washed with ether, and be dried under vacuum to obtain as light yellow solid the chloro- 3- of 5- (3a, 4,5,6,7,7a- six Hydrogen -1H- benzo [d] imidazoles -2- base) -1H- indazole NDS100281 (60mg, 40%).
1H NMR (200MHz, CD3OD) δ=8.08 (d, J=1.6Hz, 1H), 7.51 (d, J=8.8Hz, 1H), 7.23 (dd, J=2.0,8.9Hz, 1H), 3.89 (t, J=3.6Hz, 2H), 1.88-1.29 (m, 2H)；MS；275(M+H)+.
Embodiment: 4
Chloro- 1- ethyl 3a, the 7a- dihydro -1H- indazole -3- formonitrile HCN (9) of 5-:
By potassium carbonate (0.28g；1.8mmol) and bromic ether (0.2mL；1.8mmol) it is added to the chloro- 1H- indazole -3- first of 5- 4 (0.12g of nitrile；0.6mmol) in the suspension in acetone (10mL).Reaction mixture is heated 4 hours at 65 DEG C, it is cooling And it filters.Filtrate is concentrated under reduced pressure and pass through column chromatography (silica gel 100-200 mesh, 2: 8 ethyl acetate: petroleum ether) purifying with Obtain 9 (0.13g, 94%) as light yellow solid.
1H NMR (200MHz, CDCl3): δ=7.82 (t, J=1.1Hz, 1H), 7.47 (s, 2H), 4.49 (d, J= 7.2Hz, 2H), 1.57 (t, J=7.3Hz, 4H)；MS；206(M+H)+.
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- ethyl -1H- indazole (10) NDS100282:
By 9 (150mg, 0.7mmol), EDA (4mL) and P2S5The mixture of (0.065g, 0.3mmol) adds at 120 DEG C Heat 5 hours.Reaction mixture is poured into and obtains solid in ice, by filtered on buchner funnel, is washed with ether, and do under vacuum Dry (10) NDS100282 (0.11g, 60%) to obtain as pale solid.
1H NMR (400MHz, CD3OD) δ=8.12 (s, 1H), 7.68 (d, J=9.0 Hz, 1H), 7.44 (dd, J=2.0, 9.0Hz, 1H), 4.53 (q, J=7.3Hz, 2H), 3.95 (s, 4H), 1.58-1.46 (m, 3H)；13C NMR (100MHz, CD3OD) δ=159.9,138.8,130.4,128.7,127.4,122.4,119.7,111.5,44.5,13.6；MS；249(M+ H)+.
Embodiment: 5
1- benzyl -5- chloro- 3a, 7a- dihydro -1H- indazole -3- formonitrile HCN (11):
By potassium carbonate (0.35g；2.5mmol) and benzyl bromide (0.3mL；2.5mmol) it is added to the chloro- 1H- indazole -3- first of 5- 4 (0.15g of nitrile；0.8mmol) in the suspension in acetone (10mL).Reaction mixture is heated 4 hours at 65 DEG C, it is cooling And it filters.Filtrate is concentrated under reduced pressure and pass through column chromatography (silica gel 100-200 mesh, 2: 8 ethyl acetate: petroleum ether) purifying with Obtain (11) (0.18g, 80%) as light yellow solid.
1H NMR (200MHz, CDCl3): δ=7.83 (t, J=1.2Hz, 1H), 7.41-7.29 (m, 5H), 7.25-7.19 (m, 2H), 5.64 (s, 2H)；MS；268(M+H)+.
The chloro- 3- of 1- benzyl -5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole 12 (NDS100283):
By 1- benzyl -5- chloro- 3a, 7a- dihydro -1H- indazole -3- formonitrile HCN 11 (160mg, 0.6mmol), EDA (4mL) and P2S5(0.053g, 0.2mmol) is heated 5 hours under 120.Reaction mixture is poured into and obtains solid in ice, passes through Buchner funnel Filtering, washed with ether, and be dried under vacuum to obtain as pale solid 12 (NDS100283) (0.100g, 54%).
1H NMR (400MHz, CD3OD) δ=8.26-8.11 (m, 1H), 7.50 (d, J=8.8Hz, 1H), 7.34 (dd, J =1.8,8.9Hz, 1H), 7.29-7.17 (m, 5H), 4.91 (s, 2H), 3.78 (s, 4H)；13C NMR (100MHz, CD3OD)δ =160.3,139.1,136.3,134.2,128.4,127.9,127.6,127.2,127.0,122.9,120.8,111.2, 52.9 48.9；MS 311(M+H)+.
Embodiment: 6
3- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole (16) NDS100277:
According to the same procedure for synthesizing 5 (NDS100178).Yield: 51%；
1H NMR (200MHz, DMSO-d6) δ=8.25 (d, J=8.1Hz, 1H), 7.67-7.55 (m, 1H), 7.48- 7.35 (m, 1H), 7.23 (d, J=7.3Hz, 1H), 3.66 (s, 4H)；13C NMR (50MHz, DMSO-d6) δ=159.8, 141.1,135.8,126.2,122.0,121.4,121.1,110.5,49.4；MS 187(M+H)+.
1- methyl-1 H- indazole -3- formonitrile HCN 17:
According to the method for synthesis 6.Yield: 97%；
1H NMR (200MHz, CDCl3) δ=7.84 (td, J=0.9,8.2Hz, 1H), 7.57-7.47 (m, 2H), 7.37 (dd, J=4.0,8.0Hz, 1H), 4.17 (s, 3H)； MS 158(M+H)+.
Embodiment: 7
3- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl-1 H- indazole 18 (NDS100278):
According to the same procedure of 7 (NDS100179) of synthesis.Yield: 42%；
1H NMR (200MHz, CD3OD) δ=8.16 (td, J=1.0,8.2Hz, 1H), 7.57 (td, J=0.9,8.5Hz, 1H), 7.44 (ddd, J=1.0,6.9,8.4Hz, 1H), 7.37-7.20 (m, 1H), 4.22-4.06 (m, 3H), 3.78 (s, 4H)；13C NMR (100MHz, CD3OD) δ=161.0,141.1,134.1,126.6,121.9,121.8,121.5,109.3, 49.0 34.8；MS 201(M+H)+.
Embodiment: 8
3- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole (19) NDS100281:
According to the same procedure of synthesis 8.Yield: 30%；
1H NMR (200MHz, CD3OD) δ=8.09 (d, J=8.1Hz, 1H), 7.56-7.43 (m, 1H), 7.31 (s, 1H), 7.16 (d, J=7.8Hz, 1H), 3.84 (t, J=3.6Hz, 2H), 1.84-1.24 (m, 8H)；13C NMR (50MHz, CD3OD)δ =162.1,143.7,135.9,127.7,123.3,122.5,122.2,112.0,60.5,28.9,21.8；MS 241(M+H)+.
Embodiment 9:
The measurement of the insulinotropic activity of NDS100178 and NDS100179
Material and method
It is measured by as described below based on the ELISA of good optimization measurement (Shantani Protocol# 5205) The insulinotropic activity of NDS100178 and NDS100179.
Material
MIN6 cell (source: National Center for Cell Sciences, Pune)
Mouse islets element ELISA kit (Mercodia, 10-1247-01)
Plate Shaker (MixMate from Eppendorf)
HEPES balances Krebs-Ringer bicarbonate buffer (component is as follows)
* HEPES is 2- [4- (2- ethoxy) piperazine -1- base] ethanesulfonic acid
Method
I. it is testing a few days ago, the proliferating culture for being up to the MIN-6 cell that almost half converges is inoculated into 6 orifice plates In；
Ii. on the day of insulin-induced experiment, cell should become almost to converge, and be in good state.
Iii. it on the day of experiment, prepares first and contains 0.1%BSA (for rinsing) as described in material and be free of 1 × KRB-HEPES buffer of BSA (being used to balance and insulin-induced).
Iv. prepare equilibration buffer: 1 × KRB-HEPES buffer without BSA containing 3.3mM glucose is (to incite somebody to action The glucose level of all cells is reduced to phase same level).For example, adding into the 20ml KRB-HEPES buffer without BSA Add 66ul1M glucose liquid storage.
V. it prepares stimulation buffer solution (insulin-induced): (in most cases, will be containing determining glucose level 16.7mM glucose) and expectation molecular concentration 1 × KRB-HEPES buffer without BSA.
Vi. all buffers are preheated to 37 DEG C (with 15 minutes) in a water bath.
Vii. culture medium is removed from all holes, with the KRB-HEPES buffer containing 0.1%BSA of 2ml by each hole It rinses twice.
Viii. the equilibration buffer of 2ml is added in each hole, and is incubated for half an hour at 37 DEG C.
Ix. after being incubated for, the stimulation buffer solution of 2ml is added into (suitably marking) each hole, and is incubated for 1 at 37 DEG C Hour.
X. at the same time, start refrigerated centrifuge, and be 4 DEG C by temperature setting.From Mercodia ELISA kit ( Stored at 4 DEG C) in take out enzyme conjugate, dilution buffer, caliberator (Calibrator) 0 and required band (stripe) (for example, two bands for 8 samples tested in duplicate), and it is preheated to room temperature (RT).
After xi.1 hours are incubated for, the withdrawing plate from incubator does not interfere cell, and takes out the stimulation buffer solution of 1ml Into the pre-cooling pipe suitably marked.
Xii. the pipe is centrifuged 5 minutes at 600g.
Xiii. 500ul is taken out into the pre-cooling pipe just suitably marked.
Xiv. now, dilute sample (1: 30) in caliberator 0, that is, by every kind of sample of 3ul be added to 8 it is different In the caliberator 0 of 87 μ l in the 0.5ml pipe suitably marked.It is sufficiently mixed.
Xv. 1 × enzyme conjugate buffer is prepared, that is, mix the enzyme (11 ×) of 180ul with the dilution buffer of 1800ul With the ELISA for two bands.
Xvi. every part of sample (diluted, duplicate) of 10ul is added in two adjacent holes.It not touch carefully Bottom.
Xvii. the enzyme conjugate buffer (1 ×) of 100ul is carefully added into each hole, and at 25 DEG C with 800rpm is incubated for 2 hours on Plate Shaker (eppendorf MixMate).
Xviii. before the end of the incubation 45 minutes, cleaning buffer solution, substrate TMB and stop bath are taken out.They are kept At room temperature.
Xix. before the end of the incubation 5 minutes, prepare 1 × cleaning buffer solution (21 ×), that is, have in container 40ml distilled water+ 2ml (21 ×) cleaning buffer solution.
Xx. after being incubated for, each hole is cleaned 7 times with the cleaning buffer solution of 350ul.Use multiple tracks (multichannel)(175ul)。
Xxi. after cleaning, 200uL substrate TMB solution is added into each hole.Keep speed appropriate.
Xxii. make to be incubated for and carry out 15 minutes.Program is set on plate reader to read absorbance at 450nm. The shake step of addition 5 seconds is so as to be sufficiently mixed.
Xxiii. after being incubated for, the stop bath of 50ul is added into each hole with identical speed immediately, and at 450nm Read absorbance.
Determination condition is consistent in all tests.Control experiment (unused molecule processing) is in the absorbance on different dates The difference of value is attributed to cell culture condition.Other different conditions are specified in each figure.Acquired results collect as follows.
[note: if be not expressly mentioned, HG=high glucose (16.7mM), LG=low glucose (3.3mM)]
As a result
NDS100178 and NDS100179 improves the insulin secretion of glucose stimulation
In the insulin secreting ability of three as shown in figure 1 different date measurement NDS100178 and NDS100179.Following institute Show, when being handled cell 1 hour with NDS100178 or NDS100179 (the two is 33uM concentration), observes in high grape The insulin secretion of MIN6 cell significantly improves under the conditions of sugar.
The insulinotropic activity of NDS100178 and NDS100179 is glucose dependency
It was found that the insulinotropic activity of NDS100178 and NDS100179 is glucose dependency.As follows, and such as Lower glucose condition in Fig. 2 is compared, and observes pancreas in each control under the conditions of wherein cell is present in high glucose Element secretion in island significantly improves.
NDS100178 and NDS100179 show insulinotropic activity at low concentrations
As follows, when the concentration incubation with 100nM, the secretion of NDS100178 induced insulin is improved more than 100%. When concentration is reduced to 50nM or 10nM, significant difference is not obtained relative to control experiment.
It is interesting that even if NDS100179 is also active when down to the concentration of 10nM.Although in 10nM In the presence of NDS100179, do not find insulin secretion relative to the raising of control experiment be statistically significantly, but use The molecule incubation of 50nM and 100nM concentration clearly illustrates that molecule is the trend of active pancreotropic hormone agent.
The pancreotropic hormone characteristic of compound
The insulin secretion table 1 of MIN6 cell:
Compound Concentration (μM) Insulin secretion (ng/ml)
NDS100178 10.0/1.0/0.1 35.09(4.0)/12.49(1.9)/8.91(1.6)
NDS100179 10.0/1.0/0.1 15.45(1.2)/10.27(0.6)/5.55(1.0)
Value in bracket is the standard error of measured value.
Embodiment 10:
It is reduced in non-diabetic experiment mice by the glucose of NDS100179 and glucose dependent insulin is secreted
Glucose in C57BL/6J mouse reduces
By the male C 57 BL/6 J mouse overnight fasting in 12 to 13 weeks.In the morning to animal orally administration various concentration The glucose of NDS100179 or control carrier and 3g/kg.30,60,90 and after being administered before it will be administered and by compound Blood glucose is measured at 120 minutes.Reduce to molecule dose-dependant the blood glucose (Fig. 4) of C57BL/6J mouse.
Insulin in C57BL/6J mouse improves
In the research of above-mentioned progress, that is, by 12 to 13 week male C 57 BL/6 J mouse overnight fastings, and in the morning to The NDS100179 of animal orally administration various concentration or the glucose of control carrier and 3 g/kg, also measure 0 minute and 15 The insulin of minutes point.It was found that since the insulin secretion of NDS100179 is plasma concentration glucose dependency (figure 5)。
Embodiment 11:
Insulin secretion (the Glucose Stimulated Insulin of glucose stimulation is improved by NDS100179 Secretion, GSIS)
NDS100179 is the pancreotropic hormone compound that insulin secretion is only just improved under high glucose concentration.In people's pancreas The insulin measurement of NDS100179 induction has been carried out in island, mouse and MIN6 cell under the conditions of low and high glucose.It presents Data clearly illustrate that in different biosystems, it is similar (Fig. 5) that the GSIS of NDS100179 improves ability.
Embodiment 12:
Fasted plasma glucose and glucose tolerance in NDS100179- diabetes animal model (db/db mouse)
Db/db or leptin depleted mice are generally accepted diabetes models.It is given to 7 week old db/db mouse orals The NDS100179 of 10mg/kg, 30mg/kg and only carrier 30 days.Before the study began, in the 15th day and the 30th day measurement animal Fasted plasma glucose level.The significant control risen to basal plasma glucose level is shown with the animal that NDS100179 is handled, The effect depends on the amount for giving the molecule of animal.Fasting blood glucose is not shown with the animal that the NDS100179 of 30mg/kg is handled Horizontal statistically significant rising.Oral glucose tolerance test (the Oral Glucose carried out at the end of 30 days Tolerance Test, OGTT) show the significantly improving of glucose tolerance in the mouse handled through NDS100179 (Fig. 6 a and 6b)。
Embodiment 13:
Instreptozotocin Induced reservation/recovery in NDS100179- diabetes animal model (db/db mouse)
The NDS100179 for give 10mg/kg, 30mg/kg to 7 weeks db/db mouse orals and only carrier 30 days.At 30 days At the end of, assessment β cell function is calculated using HOMA- β %.The mouse handled through NDS100179 shows Instreptozotocin Induced Significantly improve (Fig. 6 c).
Embodiment 14:
The reduction of triglycerides in NDS100179- diabetes animal model (db/db mouse)
The NDS100179 for give 10mg/kg, 30mg/kg to 7 weeks db/db mouse orals and only carrier 30 days.At 30 days At the end of measure triglyceride levels.The mouse handled through NDS100179 shows the significant decrease (figure of triglyceride levels 7a)。
Embodiment 15:
There is no changes of weight in NDS100179- diabetes animal model (db/db mouse)
The NDS100179 for give 10mg/kg, 30mg/kg to 7 weeks db/db mouse orals and only carrier 30 days.Every three days Measure the weight of animals.Weight is not observed in the mouse handled through NDS100179 and has significant change (Fig. 7 b).
Embodiment 16:
The pharmacokinetic properties of NDS100179
Based on the pharmacokinetic carried out in mouse, NDS100179 is 100% orally bioavailable chemical combination Object (Fig. 9).
Measurement Value
Cmax(ng/ml) 631.1
Tmax(h) 0.25
AUC(inf)(hr*ng/ml) 854.231
AUC(0-t)(hr*ng/ml) 837.113
AUC (inf)/AUC (0-t) % 102.049
t1/2(h) 1.7
Embodiment 17:
External ADME characteristic
Embodiment 18:
The glucose uptake of NDS100179-HepG2 cell improves
NDS100179 improves the glucose uptake (Fig. 8) of HepG2 cell.Under the conditions of high glucose, trained with each drug It supports HepG2 cell 24 hours, is measured after making it have insulin resistance.It is measured after following 24 hours: cell being made to exist It is 2 hours hungry in serum free medium (containing each drug), then it is incubated for 30 minutes in KRB buffer (there is each drug), Finally it is incubated for 15 minutes with 2-NBDG (fluorescent glucose analog).Cell is sufficiently cleaned and is cracked with 1%triton-X. Fluorescence reading is read at Ex:465 and Em:540 immediately.
Embodiment 19:
The mechanism of action
NDS100178 series compound primary activation imidazoline receptor I1/I3 (IC50~50nM).In high glucose condition Under, this activation results in diglyceride (Diacylglycerol, DAG).DAG then generates arachidonic in cell Sour (Arachidonic Acids, AA).Metabolin (the especially epoxy eicosatrienoic acid of AA (epoxyeicosatrienenoic acid, EET) and hydroxyeicosatetraenoic acid (hydroxyeicosatetrenoic Acid, HETE) cause exocytosis, cause cell insulin secretion to improve.The another of glucose/DAG induction library AA can be changed One key signal transduction approach is leukotriene pathway, that is, by the way that AA to be metabolized to different leukotrienes, rather than can inhibit its pancreas The island secretory EET and HETE of element.The library AA can be maintained by the molecule that leukotriene pathway can inhibit AA metabolism, this leads to its generation It thanks only by EET and HETE, and insulin secretion therefore can be improved.Major Enzymes in leukotriene pathway first is that LTA4 is converted It is the AA derivative for inhibiting insulin secretion at the LTA4H hydrolase of LTB4.The present inventor has established a new discovery: Inhibit LTA4H that insulin secretion can be improved.NDS100178 inhibits LTA4H (IC50 < 500nM), and for coming from patent The compound 6 of US20040009976 compound 5- chloro-2-methyl -3- (4,5- dihydro -1-H- imidazoles -2- base) -1H- indoles it The IC50 of LTA4H target is greater than 10uM.
Advantages of the present invention
New antidiabetic compound.
Controlled insulin secretion relative to the glucose level in ambient enviroment.
Improve any side effect (such as hypoglycemia) of other antidiabetics.
Cardiovascular protection is provided by reducing triglycerides.
Enhancing/reservation Instreptozotocin Induced.
It is effective against 1 type and diabetes B and related complication.
Preparation method economy and commercially viable.

Claims (7)

1. the indazole compound and its stereoisomer and its officinal salt of formula 1,
Wherein the indazole compound of the formula 1 is indicated by following compounds:
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl -3a, 7a- dihydro -1H- indazole；
The chloro- 3- of 5- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole；
The chloro- 3- of 5- (4,5- dihydro -1H- imidazoles -2- base) -1- ethyl -1H- indazole；
The chloro- 3- of 1- benzyl -5- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
3- (4,5- dihydro -1H- imidazoles -2- base) -1H- indazole；
3- (4,5- dihydro -1H- imidazoles -2- base) -1- methyl-1 H- indazole；
3- (3a, 4,5,6,7,7a- hexahydro -1H- benzo [d] imidazoles -2- bases) -1H- indazole.
2. the method for being used to prepare the indazole compound of formula 1 as described in claim 1, wherein the method includes following steps It is rapid:
(i) 5- chlorisatide (1) is converted 5- chlorine indazole -3- carboxylic acid (2) by；
(ii) uses isobutyl chlorocarbonate and N-methylmorpholine to handle the compound 5- chlorine obtained in step (i) under argon gas Indazole -3- carboxylic acid (2) is then reacted with ammonia water to obtain the chloro- 1H- indazole -3- formamide (3) of 5-；
(iii) the chloro- 1H- indazole -3- formyl of compound 5- that pyridine and trifluoroacetic anhydride processing obtain in step (ii) Amine (3) is to obtain cyano compound (4)
(iv) makes the cyano compound (4) obtained in step (iii) and potassium carbonate and alkyl halide or benzyl bromide in acetone Reaction be substituted indazole formonitrile HCN (6), (9), (11), (17), and in P2S5In the presence of make the indazole formonitrile HCN being substituted (6), (9), (11), (17) and diamine reactant are to obtain 1 compound of formula described in claim 1
Or
In P2S5In the presence of make the cyano compound (4) obtained in the step (iii) and diamine reactant to obtain claim 1 1 compound of formula.
3. method according to claim 2, wherein the alkyl halide used in step (iv) is selected from bromic ether and methyl Iodine.
4. method according to claim 2, wherein the diamines used in step (iv) is selected from 1,2- cyclohexane diamine And ethylenediamine.
5. pharmaceutical preparation, it includes as 1 compound of formula as described in claim 1 of active constituent or its stereoisomer, And its officinal salt, and one or more of pharmaceutical acceptable carrier, diluent or excipient.
6. the indazole compound of formula 1 as described in claim 1 is used to prepare for treating diabetes, diabetic complication, generation It thanks disorder, cardiovascular functional disorder or in which there are glucose processing to be damaged, triglyceride levels change or Instreptozotocin Induced drop The purposes of the drug of low related disease.
7. purposes as claimed in claim 6, wherein the drug be configured to for 1 compound of formula of therapeutically effective amount or its Stereoisomer and its officinal salt are applied to mammal.
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